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3, 3′,5 ,5′-Tetramethylbenzidine Liquid Substrate, Supersensitive, for ELISA

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aqueous solution

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shipped in

wet ice

shipped in

wet ice

shipped in


shipped in

wet ice

storage temp.


storage temp.


storage temp.


storage temp.


General description

This product contains 3, 3′,5 ,5′-Tetramethylbenzidine (TMB) in a mildly acidic buffer. The substrate is supplied as a one component ready to use solution. Rate kinetics are approx. 40% faster than traditional TMB formulations. Not recommended for membrane or immunohistochemical applications that require a precipitating reaction product.


3, 3′,5 ,5′-Tetramethylbenzidine Liquid Substrate, Supersensitive, for ELISA has been used as a substrate for peroxidase:
  • to determine synthesized hyaluronan and HAS2 (hyaluronan-synthesizing enzyme) expression by the transfectant
  • to measure the levels of α-hemolysin toxin
  • in imaging bound IgGs

3,3′,5,5′-Tetramethylbenzidine (TMB) is a peroxidase substrate suitable for use in ELISA procedures. The substrate produces a soluble end product that is pale blue in color and can be read spectrophotometrically at 370 or 655 nm. The TMB reaction may be stopped with 2 M H2SO4 (resulting in a yellow color), and read at 450 nm.

Storage Class Code

10 - Combustible liquids



Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

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The activity of hyaluronan synthase 2 is regulated by dimerization and ubiquitination
Karousou E, et al.
The Journal of Biological Chemistry, 285(31), 23647-23654 (2010)
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Nitroblue Tetrazolium (NBT) is used with the alkaline phosphatase substrate 5-Bromo- 4-Chloro-3-Indolyl Phosphate (BCIP) in western blotting and immunohistological staining procedures. These substrate systems produce an insoluble NBT diformazan end product that is blue to purple in color and can be observed visually.

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